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目的观察经过结构改建的丁型肝炎病毒核酶（ＨＤＶ核酶）对丙型肝炎病毒（ＨＣＶ）基因组ＲＮＡ是否存在剪切作用。方法对ＨＤＶ基因组核酶的茎ＩＶ区和底物结合区进行改建，获得３种针对ＨＣＶ ＲＮＡ的ＨＤＶ核酶ＲｚＣ１、 ＲｚＣ２和ＲｚＣ３。体外转录制备含ＨＣＶ ＲＮＡ ５’一非编码区（５’－ｎｏｎｃｏｄｉｎｇ ｒｅｇｉｏｎ， ５’－ＮＣＲ）及部分 Ｃ区的底物 ＲＮＡ（ＨＣＶ ＲＮＡ ５’－ＮＣＲ－Ｃ），进行 ５’端放射性标记。在一定的 ｐＨ、温度、 Ｍｇ２＋浓度和有或无去离子甲酰胺存在等条件下，将核酶和底物按摩尔比１００：１混合，反应２ ｈ后聚丙烯酰胺凝胶电泳，放射自显影，推算剪切百分率。结果 ＲｚＣ１和ＲｚＣ２可剪切ＨＣＶ ＲＮＡ ５’－ＮＣＲ—Ｃ，在适宜浓度的去离子甲酰胺存在时剪切效率较高。ＲｚＣ３对底物几乎无剪切活性。结论设计得当的ＨＤＶ基因组核酶可以剪切异源性ＲＮＡ分子ＨＣＶ ＲＮＡ。
Objective To investigate whether the structurally modified hepatitis D virus ribozyme (HDV ribozyme) is capable of cleaving the hepatitis C virus (HCV) genomic RNA. Methods The stem IV region and substrate binding region of HDV genomic ribozyme were reconstructed to obtain three HDV ribozymes RzC1, RzC2 and RzC3 targeting HCV RNA. Substrate RNA (HCV RNA 5’-NCR-C) containing 5’-noncoding region (5’-NCR) and part C region of HCV RNA was prepared by in vitro transcription and radiolabeled at the 5 ’end. Under certain conditions of pH, temperature, Mg 2+ concentration and the presence or absence of de-ionized formamide, the ribozyme and substrate were mixed at a molar ratio of 100: 1. After reaction for 2 h, polyacrylamide gel electrophoresis and autoradiography , Calculate the cut percentage. Results RzC1 and RzC2 cleaved the HCV RNA 5’-NCR-C, resulting in higher shearing efficiency in the presence of deionized formamide at a suitable concentration. RzC3 has almost no cleavage activity on the substrate. Conclusion The well-designed HDV genomic ribozyme can cleave the heterologous RNA molecule HCV RNA.